title: Detection of chitinase activity by 2-aminobenzoic acid labeling of chito-oligosaccharides.
abstract: Chitinases are hydrolases capable of hydrolyzing the abundant natural polysaccharide chitin. Next to artificial fluorescent substrates, more physiological chito-oligomers are commonly used in chitinase assays. Analysis of chito-oligosaccharides products is generally accomplished by UV detection. However, the relatively poor sensitivity poses a serious limitation. Here we report on a novel, much more sensitive assay for the detection of chito-oligosaccharide reaction products released by chitinases, based on fluorescent detection, following chemical labeling by 2-aminobenzoic acid. Comparison with existing UV-based assays, shows that the novel assay offers the same advantages yet allows detection of chito-oligosaccharides in the low picomolar range.
authors.affiliation: Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Karen
authors.initials: K
authors.lastname: Ghauharali-van der Vlugt
authors.forename: Anton P
authors.initials: AP
authors.lastname: Bussink
authors.forename: Johanna E M
authors.initials: JE
authors.lastname: Groener
authors.forename: Rolf G
authors.initials: RG
authors.lastname: Boot
authors.forename: Johannes M F G
authors.initials: JM
authors.lastname: Aerts
mesh_terms: D001681:Biological Assay; D002686:Chitin / Q000032:analysis / Q000737:chemistry*; D002688:Chitinases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry*; D013379:Substrate Specificity; D062367:ortho-Aminobenzoates / Q000737:chemistry*
chemical_list: D009844:Oligosaccharides; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D002686:Chitin; D002688:Chitinases